1. Pedigree ALS85 M M M M W W W W M M W W W W W Familial ALS M377V Sporadic ALS G294A Sporadic ALS Q331K RNA binding proteins and neurodegeneration: TDP 43 Sreedharan et al. Science 2008 Wild type M337V Chick embryo Developmental delay Transverse Section IHC Transgene expression Tunel Staining Apoptotic Cells FTD ALS/MND

2. Average age at onset 35 years Average survival 12 months Linkage to Chromosome 16 LOD score = 4 Ruddy et al. AJHG 2003 RNA binding proteins and neurodegeneration: FUS Arg 515 Gly Arg 521 His Arg 521 Cys 3 different missense mutations 8 unrelated familial ALS kindreds FUS R514G Wildtyp e FUS GFP-FUSAnti-FUSDAPIMerge Mutations cause FUS to relocate to the cytoplasm Because they disrupt their nuclear localising signal Mutant FUS forms cytoplasmic inclusions In ALS patients Vance et al. Science 2009

3. Toxic RNA expansion and neurodegeneration: ORF72 = ALS = FTD = ALS-FTD Multigenerational Dutch kindred ALS and FTD variable phenotype Linkage to Chr 9p LOD score = 3 Vance et al. Brain 2006 Le Ber et.al 2009 Gijselinck 2010 Momeni et.al 2006 Morita et.al 2006 Vance et.al 2006 Valdmanis et.al 2007 Pearson 2011 Boxer et.al 2010 ~ 4MB Overlap 10 kindreds linked Region narrowed to 4MB BAC haploid clones DNA capture Brain RNA sequenced Whole genome Next generation seq. 97 SNP common haplotype C9ORF72 intronic hexanucleotide (GGGGCC)n expansion mutation 700 to 1,000 repeats De Jesus-Hernandez Neuron 2011 Renton et al Neuron 2011 Expanded Intronic repeat detected by PCR and Southern Blot